Cloning and expression of the pneumococcal neuraminidase gene in Escherichia coli

Anne M. Berry; James C. Paton; Eric M. Glare; David Hansman; David E.A. Catcheside

doi:10.1016/0378-1119(88)90046-7

Cloning and expression of the pneumococcal neuraminidase gene in Escherichia coli

Anne M. Berry, James C. Paton, Eric M. Glare, David Hansman, David E.A. Catcheside

Research output: Contribution to journal › Article › peer-review

24 Citations (Scopus)

Abstract

A gene bank of Sau3AI-generated Streptococcus pneumoniae DNA fragments was constructed in Escherichia coli K-12 by cloning into the BamHI site of the cosmid vector pHC79. One clone capable of cleaving the fluorogenic neuraminidase substrate 2'-(4-methylumbelliferyl)-α-d-N-acetyl-neuraminic acid was isolated. This activity was inhibited by treatment with a mouse antiserum raised against purified pneumococcal neuraminidase. The recombinant plasmid purified from this clone (designated pJCP301) contained approximately 3.0 kb of pneumococcal DNA. Western-blot analysis indicated that E. coli K-12[pJCP301] produced a 98-kDa polypeptide which reacted with antineuraminidase serum.

Original language	English
Pages (from-to)	299-305
Number of pages	7
Journal	Gene
Volume	71
Issue number	2
DOIs	https://doi.org/10.1016/0378-1119(88)90046-7
Publication status	Published - 30 Nov 1988

Keywords

cosmid vector
recombinant DNA
Streptococcus pneumoniae
virulence factor, gene bank

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title = "Cloning and expression of the pneumococcal neuraminidase gene in Escherichia coli",

abstract = "A gene bank of Sau3AI-generated Streptococcus pneumoniae DNA fragments was constructed in Escherichia coli K-12 by cloning into the BamHI site of the cosmid vector pHC79. One clone capable of cleaving the fluorogenic neuraminidase substrate 2'-(4-methylumbelliferyl)-α-d-N-acetyl-neuraminic acid was isolated. This activity was inhibited by treatment with a mouse antiserum raised against purified pneumococcal neuraminidase. The recombinant plasmid purified from this clone (designated pJCP301) contained approximately 3.0 kb of pneumococcal DNA. Western-blot analysis indicated that E. coli K-12[pJCP301] produced a 98-kDa polypeptide which reacted with antineuraminidase serum.",

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AU - Berry, Anne M.

AU - Paton, James C.

AU - Glare, Eric M.

AU - Hansman, David

AU - Catcheside, David E.A.

PY - 1988/11/30

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AB - A gene bank of Sau3AI-generated Streptococcus pneumoniae DNA fragments was constructed in Escherichia coli K-12 by cloning into the BamHI site of the cosmid vector pHC79. One clone capable of cleaving the fluorogenic neuraminidase substrate 2'-(4-methylumbelliferyl)-α-d-N-acetyl-neuraminic acid was isolated. This activity was inhibited by treatment with a mouse antiserum raised against purified pneumococcal neuraminidase. The recombinant plasmid purified from this clone (designated pJCP301) contained approximately 3.0 kb of pneumococcal DNA. Western-blot analysis indicated that E. coli K-12[pJCP301] produced a 98-kDa polypeptide which reacted with antineuraminidase serum.

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KW - recombinant DNA

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KW - virulence factor, gene bank

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Cloning and expression of the pneumococcal neuraminidase gene in Escherichia coli

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